Areas of Legal Liability for Advanced Practice Nurses Essay

Areas of Legal Liability for Advanced Practice Nurses - Essay Example The failures to follow standards of care may include failure to institute a protocol or failure to use proper procedure for a particular skill. Under such circumstances, nurses are liable for damages caused as a result of their failure to implement proper actions. Hence, management should be ready to take necessary actions against nurses who violated proper standards of care. Such a response may assist the management to limit the liability for managers and the facility. Failure to use equipment responsibly falls under the category of direct breach of care, standards and it can be more easily proved. If a nurse deploys a clinical equipment for any use other than it was actually intended, he/she is liable to face legal consequences. In addition, it is observed that nurses often use equipments that they have not been adequately trained to use. If the misuse of the equipment causes any harm to patients, the nurse is in legal jeopardy. Hence, the management must ensure that they have employed adequate number of skilled and experienced nursing staff to prevent nurses from risking the life of patients. An effective nurse-patient ratio would reduce equipment errors significantly. In most of the nurse malpractice suits, an element of poor communication between nurse and patient, nurse and physician, or nurse and other healthcare providers is identified. The communication failure cases may include nurses’ failure to communicate all relevant patient data to the physician or discharge information to the patient. In order to avoid such troubles, the management may insist nurses to record all matters concerning patient care appropriately. In addition, it is advisable for the management to establish an effective reporting system in the care facility so that the flow of communication between nurses and other parties would become effective. In the eyes of law, if a nurse fails to document something appropriately, the court will consider as it

Personal statement for ucas Example | Topics and Well Written Essays - 500 words

For ucas - Personal Statement Example I have practiced dancing for twelve years and instructed dancing in schools like IISC. Through my dancing skills, I have managed to achieve a government award in 2007. I love concerts and seldom miss any occasion. I have participated in cultural events like at the Indus, and danced at the Indus cultural festival like the ‘Mamma Mia, and We Will Rock You Musical’. As a kid, I had the passion of becoming an engineer, which motivated me to excel in subjects like Mathematics and Physic. I have conducted a full length research on, "How the temperature affects the magnetic field of different magnets" and wrote a 4000 words essay on the research. I conducted this research in a PHD research university in my 11th grade summer vacations. As a young adult, I had asked for career advice from a motivational speaker who had visited my school. Henceforth, I have found learning Mathematic to be interesting, since it is a powerful tool for analyzing and solving problems. Mathematics has several specialization segments like pure and applied mathematics. Additionally, I love Physics since it assists in understanding the world deeply by its information. I learned about Electrical Engineering in my initial school and assistance from my bigger brother. He is a Civil Engineer doing well in his line of specialty. Furthermore, he is my role model, since we share the same dream of being engineers. In particular, I realized that my passion was in Electrical Engineering. My career master rightly advised me on the path to follow to be an Electrical Engineer, which I followed it to the latter. My college years have vastly acquired me with educational and managerial skills. My internship at the Battery Industry (Amaron Battery Industry) instilled managerial and leadership skills in me. I was exposed to dealings within the learning institution; hence I am armed with administration and managerial skills. Taking part in the science

Meaning of Life and Success Essay Example for Free

Meaning of Life and Success Essay Success is a person or thing that desired aims and achieves or attains fame or any form of accomplishment which will always make them proud or happy at the end of either the day, month or year. The word success was originated in the mid-16th century from a Latin word successus and a verb succedere meaning come close after. When I hear of the word success there are two things that come in mind, hard work and determination. Hard work is one of the main and most important building blocks in my success foundation. Working hard is one of the best and most accurate ways to think about being successful. Success cannot be earned unless you strive and work hard for it, because it is a word that comes with work and if work is not done then the meaning of the word can’t be accomplished. Like Einstein said â€Å"If A is a success in life then A equals x plus y plus z. work is x; y is play and z is keeping your mouth shut† for instance to me as an athlete working hard and in playing hard rather than running my mouth are the best way of being successful in game point stands in my career. Determination is a very critical and important aspect when it comes to success. To be determined to achieve any goals or aims is a very important choice of being successful. Bertrand 2 Determination can to be physically, spiritually and emotionally which all combines with hard work and if you combine both hard work and determination together you will always get a good result of success. Being successful in life can be one of the precious and happiest moments in life that can last for a long time if and only if you do what is expected of you to keep the success going on. Success can be advantageous in many ways; Accomplishing a goal, moving on to a new goal, brings happiness and fulfillment, set new and personal records, inspire, motivate and give you strengths to aim high and set bigger goals. But it’s not always everybody that’s successful that is happy or satisfied with their success. The isn’t that much difference being successful than being a failure, like the good old quote that say â€Å"the difference between a successful person and others is not a lack of strength not a lack of knowledge, but rather the a lack in will† this quote is very simple and well understood and the quote’s point is clearly pointed out, which state the main difference between a successful person and others which is the will to be the successful person that they are. To be successful person there should be some sought of guidelines to help you achieve or attain success. Thinking about success also brings about the thought or idea of failure â€Å"In order to success your desire for success should be greater rather than your fear of failure†. An important and inspiring quote as this from one of the favorite, idol and inspiring actor in the movie industry and in the society should and is been considered by a lot of fans to be very important in achieving success rather than having the fear of failure because his path of success has given or got him to where he is at the moment.

Seeing As Being Prepared To See Philosophy Essay

Seeing As Being Prepared To See Philosophy Essay Ralph Waldo Emerson says aptly: People only see what they are prepared to see  [1]  . It means that people will only see thing as they want it to be. On the other hand, it simply means that we see things as we are. Why could not we see things as they are instead as we are? Therefore, how can we be sure that what we perceive now is the way it is supposed to be? The reasons of why this happened because of the ways of knowing. There are four ways of knowing that can misled our seeing and understanding of the things which are perception, reason, emotion and language. But however without them, we cannot create knowledge of reality and truth because brain does not have a direct contact to the real world. It is somehow these ways of knowing do help us to see and understand things as they are but just to a certain extent. Therefore, in this essay, I intend to discuss to what extent we see and understand things not as they are but as we are. Language is a conventional code of symbols that allows a sender to formulate a message that can be understood by a receiver. How we see things is strongly influenced by our language and our seeing also makes influence on our thinking. Therefore, our thinking cannot be separated from our language and even we could say that our language limits our thinking. According to the Linguistic Relativity Theory, an individuals nature language determines the way the individual thinks and perceives the world which also can bewitch the intelligences  [2]  . One example is infinite monkey theorem. This theorem states that a monkey hitting keys at random on a typewriter keyboard for an infinite amount of time will almost surely type a particularly chosen text, such as the complete works of William Shakespeare  [3]  . People always misinterpret by the true meaning of this theorem (by linguistic and perception). With the picture of the chimpanzee typing a typewriter will cause people to regard and value it as an art. People may have thought that the picture of the chimpanzee is the same as the Cassius Marcellus Coolidges work of his paintings in the dogs playing poker  [4]  genre. But the truth is the picture and the theorem is all about Mathematics that illustrates the perils of reasoning about infinity by thinking a vast but finite number and vice versa From the context, the words of almost surely is a mathematical term with precise meaning and the monkey is not an actual monkey but it is a metaphor for an abstract devices that produces a random sequences of letters ad infinitum. At first we really do see it not as the theorem but instead as the art of the monkey. It is because the sense of our sight which is perception and the language gi ve us false idea of what the theorem really is. Hence, the language itself will limit our seeing and understanding things not as they are but as we are. Unless we already study about the theorem beforehand, we will know what the picture of chimpanzee and the context of the sentence of the theorem are trying to convey the meaning. So, we will see the theorem as it is but not as we are. This implies that many words have no true meaning; rather they have so many different meanings which can only be appreciated in context. Therefore, we must be aware of the true meaning to be able to use a word accurately because word can mean so many things in so many situations that require us to perceive it based on our knowledge and experiences which are often being limited by our senses. So, one must understand the context, or background, in which a word is used to have a grasp on the meaning of the word itself. Understanding the context of a word is nearly as important as an understanding of the word itself, as the situation controls to a degree how the word will be used.The result would be language which is far more clear, precise, and less mislea ding, or bewitching. When language free of most problems it would make it an even greater tool and developing better understanding and knowledge through this communication, ultimately it would help us to see things as they are. Moving on to science, I believe there is always a new paradigm to it due to scientists see things as (we are) where suppose they should have see things as they are. Why does paradigm changes from time to time? Does paradigm occur because of we (scientists) see and understand things not as they are but as we are (scientists)? According to the historian of science, Thomas Kuhn, paradigm is the word refers to the set of practices that defines a scientific discipline at any particular period of time.  [5]  In other words, scientists have always work based on their paradigm which is a normal science of that particular scientific community. Normal science is an assumption (might be deceived by the perception, emotion and reasoning) that the scientific community knows what the world is like. So, scientists will adjust and modify their paradigm if falsifications become apparent but consistently stay within it. Eventually, there comes a point when new observations are no longer compatible with the existing paradigms. From here the revolution occurs and new paradigm will replace the old one. All this is happening because the paradigm itself is a human construct and all the scientific observations are made by using our human senses, human intelligences and human rationality which the ways of knowing are necessary in these processes. However, these ways of knowing (perception, emotion, language and reasoning) that exist among scientists can limit their capabilities to see things as they are. Therefore, scientists will always come about with new ideas, assumptions and theory that cause the amendments of the paradigm. To further up, according to Kuhns book, The Structure of Scientific Revolutions  [6]  , he said that the perception of the world depends on how the percipient conceives the world where two scientists who witness the same phenomenon and are steeped in two radically different theories will see two different things. One of the examples is the ideas of the Charles Darwin and Abbot Gregor Johann Mendel about the inherited characteristics from two parents into their child  [7]  . Darwin suggested that the characteristics of the mother and father were blended to produce a child who looks similar to both. Abbot Gregor Johann Mendel developed theories over seven years by studying and testing pea plants. In the 1930s, the Mendels conjectures, The Law of Segregation and the Law of Independent Assortment were found correct after the genetics and research into inheriting traits began to be investigated. On the other hand, Darwins speculations of the blending theory only pervaded into the first offspring of two parents but not with the characteristics which Darwin could not explain, but Mendel did. This shows that the two scientists have two different theories on the same phenomenon because of the perception, emotion and reasoning are different to each other. But this paradigm could not be a promising in the future since paradigm always changing based on human being observations and assumptions that are mainly seize by our ways of knowing. The first principle is that you must not fool yourself and you are the easiest person to fool.   (Richard Feynman, American theoretical physicist, 1918-1988)  [8]   Even though sciences always give us the areas of uncertainty, but without sciences we would not be able to know the world. We could not see the things as they are without the existence of science. Whatever inadequacies as a way-of-knowing science may have are inadequacies caused by the fact that it is a human construct but there is no way-of-knowing created by humans will ever be entirely reliable, entirely precise, and entirely objective. The way we develop our scientific knowledge, science as a way-of-knowing is pragmatic. Thus, it must be consider as reliable, precise and objective. On the other hand, there is a scientist who models their claim on science for good reason. It also can be the most reliable way-of-knowing and be the best justified true belief if we are limiting ours way-of-knowing to the physical and world around us. Without our realisation, there is an absolute way-of-knowing in which justification is absolutely independent of observation. Plus, there is also an obs ervation that requires our justification that based on our way-of-knowing solely. Hence, the thing that we see and understand may do not need us to see them as they are but as we are. In conclusion, we do see things as we are but not as they are but just to a certain extent. All the areas of knowledge will help us to see and understand things more as they are but not as we are. Although there is some part that we as a human are not capable of seeing and understanding the thing as they are since our ways of knowledge can be deceiving but we can be guided by any theories in Mathematics and Sciences. Not only that, with the developing technologies we will eventually see and understanding things as they are and we can reassure our belief in the world.

Strains of ESBL Producing E. Coli | Investigation

Strains of ESBL Producing E. Coli | Investigation Introduction Background of Study Extended Spectrum Beta- Lactamases (ESBL) are beta lactamases which are mainly produced by family members of Enterobacteriaceae derived from mutations of the previous broad-spectrum beta-lactamase (Sharma et al., 2010). This enzyme works by hydrolysing and destroying the ÃŽ ²- Lactam ring of all cephalosporins, penicillins and monobactams (Sharma et al., 2010). In recent years, the emergence of ESBL producing Escherichia coli has posed a very serious problem to the management of diseases caused by this organism as only limited choice antibiotics can be given to patients. Carbapenems are the drugs of choice for the treatment of ESBL producing E.coli, however, carbapenamase resistance has recently been reported (Paterson and Bonomo, 2005). Prolonged use of antibiotics was suggested as the main cause of the emergence of ESBL E.coli and the fact that the genes coding for ESBLs are easily transferred from one organism to another organism via conjugation, transduction and transformation ma ke the spread even quicker (Vaidya et al., 2011). ESBL producing organisms were first reported from a patient in Germany in 1983 and since then , several outbreaks have been reported worldwide usually one particular â€Å"super† strain has been involved presumably combining not only the capability to produce ESBLs but also possessing various other virulence factors that contribute to their pathogenic success. (Harada et al., 2013). These pathogenic ESBL producing Escherichia coli in recent years have become a major concern and their emergence is now become alarming in clinical fields and subjected to comprehensive studies worldwide. The most common infections caused by pathogenic ESBL producing E.coli are urinary tract infections (UTI), bloodstream infections, gastrointestinal infections (Fatima et al., 2012; Bekat et al., 2002). According to Petty et al., (2013), globally, E.coli sequence type ST131 is the multidrug resistant clone strain which is responsible for ESBL CTX-M15 bearing genes, and it is the most alarming pathogenic ESBL producing E.coli associated in causing UTIs and septicaemia in hospital community acquired infections. ? in UK or worldwide? As genes coding for ESBL in Escherichia coli are known to be transferable this raises further fear of the spread of these genes to other strains, continuous monitoring of the predominant strains of E.coli which carry the ESBL genes is therefore important. Problem statement Studies of ESBL producing Escherichia coli in the South Manchester population have been carried out previously. This study will investigate strains of ESBL producing E. coli currently circulating in the Stockport Population of South Manchester and compare them to those delineated in the previous studies using a molecular typing and pulse-field gel electrophoresis. Objectives The objectives of the project are: Screen for ESBL Escherichia coli clinical isolates Identify strain using PFGE Assess the relatedness of the strains by PFGE analysis Determine Escherichia coli plasmid profile Identify Escherichia coli phylotyping group 1.0.4. Significance of study Finding from this study will contribute to the existing data and the body of knowledge on the molecular relationship of predominating of E.coli isolates from South Manchester populations. 1.0.5. Scope and Limitations There are no data on the antibiotics consumed by the patients in which the clinical isolates originates from. The availability of this data might help in understanding relationship between an exposures of certain antibiotics to the emergence of ESBL producing E.coli strain. PFGE also has several limitations in which the method assess visual relatedness of an isolates and not using a phylogeny relationship which provide more accurate molecular relationship between an isolates. Escherichia coli Escherichia coli is a motile gram negative rod, facultative anaerobe, non- spore forming bacteria taxonomically belong to the family of Enterobacteriaceae. It is considered as a normal inhabitants of gut and intestine in almost all warm blooded mammals and found as a faecal contaminant in the environment (Brennan et al., 2010; Darnton et al., 2007; Diniz et al., 2005). Most varieties of E.coli are harmless and do in the most part contribute to the normal and healthy intestine condition, while a few cause limiting abdominal cramp associated with diarrhoea. However, there are some serotypes that becoming a major threat to the human health, because they have acquired certain genetic material and virulence factors which enabling them transformed into pathogenic E.coli causing broad spectrum of disease (Clarke et al., 2003; Kaper et al., 2004). Pathotypes of E.coli are classified by specific mechanism in which they causing a disease, presence of certain virulence genes and their clinical manifestations (Chang et al., 2004). Growth requirements E.coli are non- fastidious bacteria, thus it can be cultured in artificial media with various altered physical and nutritional growth factors. It can be isolated easily from clinical samples by culturing into culture media and incubated at optimum temperature of 37 ºC anaerobically or aerobically as it is a facultative organisms (Yunlin et al., 2004) Uropathogenic Escherichia coli According to Pitout et al., (2005) E. coli is a frequent cause of the urinary tract infections (UTIs) of a hospitalised and non- hospitalised patients. UTIs are usually self- limiting but untreated lower urinary tract infections such as simple cystitis (bladder infection) can lead to much more severe illness known as pyelonephritis (renal infections) mainly among adult women (James et al., 2011). Infections occur by ascending movement of E. coli up the periurethral area colonising the bladder or infections by movement down from the intestinal tract. Due to anatomical complexities in women, they are more prone to be diagnosed with UTIs for at least once in their lifetime (James et al., 2001) 1.3  Escherichia coli typing 1.3.1  Plasmid profiling Multidrug resistant bacteria including ESBL producing Escherichia coli acquire their resistance by various gene transfer mechanisms which include transformation, horizontal transfer either by transduction, and conjugation, transposon and most often, are plasmid mediated (Carattoli et al., 2005) Plasmids are an extra chromosomal fragments of self- replicating DNA present in most of the bacterial species. Plasmids contain genes that are an essential for the replication of genes that promotes resistance to agents such as antibiotics, ultraviolet radiation, metals and bacteriophages. 1.3.2  Pulse-field gel electrophoresis PFGE was developed and described first by Schwartz and Cantor (1984). It is a molecular technique of typing a bacteria especially pathogenic Escherichia coli 0157:H7, non 0157: H7, Salmonella serotypes, Shigella sonnei and Shigella flexneri. PFGE uses a gel electrophoresis- based technique that allows separation of large molecular weight DNA up to 2Mb- 10Mb using a standard PFGE method (CDC, 2013; Hansen et al., 2002; Vimonet et al., 2008) PFGE is different to conventional gel electrophoresis as the large genomic DNA is digested with restriction enzyme that recognise and cleave specific sequences of DNA known as restriction site in an organism to produce a multiple DNA fragments which differ in size of their molecular weight (Van der Ploeg et al., 1984). The fragments are then run through constant changing electric field of PFGE resulting in a formation of DNA at various discrete size bands. This typing method has also been shown to have more discriminating power and reproducibility between laboratories than the newer molecular typing method such as ribotyping and multi- locus sequence typing (MLST) which confer more on the global epidemiology and revolutionary relationship between bacterial species (Vimonet et al., 2008) 1.3.3.  Escherichia coli phylogenetic group 2.0  Materials and Methods 2.0.1  Bacterial Isolates Bacterial isolates used in this study were Escherichia coli clinical isolates which was collected from Stepping Hill Hospital. Isolates undergo an anonymisation numbering of 1 to 20. 2.0.2.  Bacteriological Media The media used in the study were a selective differential medium for UTI Escherichia coli which is Chromogenic agar and nutrient agar which was used as a medium for growth and maintenance of isolates. 2.0.3  Antibiotic disks Table 1: Antibiotic disks used in this study was obtained from Oxoid.Ltd. Antibiotics Antibiotic Group Gentamicin (10 µg) Aminoglycosides Ciprofloxacin (5 µg) Quinolone Amoxicillin (25 µg) Penicillin Cefpodozime (10 µg) Cephalosporin Mecillinam (10 µg) Beta lactam Trimetophrim (2.5 µg) Bacteriostatic ESBL Disk kit (Mast Diagnostics) 2.0.4  Buffers and solutions Tris Borate EDTA (TBE X1 and X0.5) (Sigma) pH 8.2 was used as a running buffer in agarose gel electrophoresis. 2.0.5  Commercial kits The commercial kit used in this study was QIAprep Spin Miniprep Kit (Qiagen) and DNeasy Blood and Tissue Kit (Qiagen) 2.1.  Screening for multidrug resistance and potential ESBL producers in Escherichia coli clinical isolates Antibiotic susceptibility of Escherichia coli to six antibiotics (Table 1) were tested using the Kirby Bauer disk diffusion method. A 24 hour cultures from Nutrient agar was used. Then, a single colony was taken and transferred into 5ml Mueller Hinton Broth. It was then incubated at 37 °C to develop a heavy suspension of overnight cultures. A sterile cotton swabs were used to streak onto the Mueller Hinton agar and the rotation were repeated for three times. A final sweep was made around the rim of the agar. The plates were allowed to dry for several minutes. Using antibiotic dispenser, the disk that has been impregnated with a fixed antibiotic concentration was placed on the surface of the agar surface. After 24hr of an incubation period, the plates were checked for the presence of inhibition zone. Each recorded inhibition zone was compared with antimicrobial susceptibility testing disc chart provided by The British Society for Antimicrobial Chemotherapy (BSAC). The inhibition zon e of each antibiotic was reported as ‘sensitive’, ‘intermediate’ or ‘resistance’. Isolates showing resistance to three or more classes of antibiotics were considered as multidrug resistance (Falagas, 2007). ESBL producers were detected by testing sensitivity of isolates against a pair discs (cefpodoxime 10 µg and cefepime 10 µg) with and without clavulanic acid placed oppositely on an agar. According to manufacturer (Mast diagnostics), isolates were considered as an ESBL if there is a presence of 5mm larger inhibition zone in disks with clavulanic acid rather than the disks without the clavulanic acid. 2.2. Determination of plasmid profiles in MDR and ESBL Escherichia coli 2.2.1  Plasmid Extraction Prior to Plasmid DNA extraction, a fresh overnight cultures of E.coli after an incubation at 37 ºC in a Mueller Hinton broth were harvested. Plasmid DNA extraction was carried out using QIAprep Spin Miniprep Kit (Qiagen) following the manufacturer’s instructions. Extracted plasmid DNA was stored at -20 ºC until use. 2.2.2  Detection of plasmid by agarose gel electrophoresis The profiles of the plasmid DNA was determined on a 0.7% agarose gel electrophoresis which has been carried out at 70 Vcm-1 for 120 minutes. The size of DNA bands was estimated using Hyper ladder 1 (Bioline) as a reference molecular weights marker. The bands were visualized under UV transilluminator and photographed with digital camera connected to visualisation unit (Alpha Innotech) and the size of the plasmid were measured by visual comparison to the reference marker. 2.3  Escherichia coli pathotypes determination 2.3.1.  Genomic DNA extraction Primary cultures on the nutrient agar was inoculated into 3ml Mueller Hinton broth for 24 hours at 37 ºC. The cells was then harvested by centrifugation at 12, 000 for 3 minutes. Genomic DNA extraction was carried out using DNeasy Blood and Tissue (Qiagen) kit following the manufacturer’s instructions. Final volume of 150 µl genomic DNA were collected and kept at -20 ºC until needed. 2.3.2  Multiplex PCR for Escherichia coli phylotyping PCR reaction mix preparation must be carried out on ice. PCR was performed in 0.2ml PCR tubes on a GeneAmp PCR System 9700 thermocycler (Applied Biosystems ®) with a total 25 µl of reaction volume as described in Table 2 and PCR condition according to Table 3. The negative control reaction lacking the DNA was included. Table 2:  PCR reaction mix Components Required concentrations Volume ( µl) per reaction Biomix Red 2X 12.5 Primer (forward) chuA yjaA tspE4.c2 20pmol 20pmol 20pmol 1 1 1 Primer (reverse) chuA yjaA tspE4.c2 20pmol 20pmol 20pmol 1 1 1 DNA 2 Ultrapure sterile water 4.5 Total volume per reaction 25 Table 3: Conditions for PCR gene amplification Genes Primer sequence PCR condition chuA Forward 5’-GACGAACCAACGGTCAGGAT-3’ Reverse 5’-TGCCGCCAGTACCAAAGACA-3’ Initial denaturation: 94 °C for 4 mins Denaturation: 94 °C for 25 secs Annealing: 52 °C for 40 secs 30 cycles Extension: 72 °C for 50sec Final extension: 72 °C for 6 mins yjaA Forward 5’-TGAAGTGTCAGGAGACGCTG-3’ Reverse 5’-ATGGAGAATCGGTTCCTCAAC-3’ tspE4.c2 Forward 5’-GAGTAATGTCGGGGCATTCA-3’ Reverse 5’-CGCGCCAACAAAGTATTACG-3’ 2.3.3  Detection of by agarose gel electrophoresis After completion of the multiplex PCR, the amplification product were separated by dry electrophoresis system. 15 µl of amplified product was mixed with 5 µ 5X DNA loading buffer (Bioline) and loaded onto 2% agarose gel incorporated with SYBR green dye. After electrophoresis, the gel was visualised by exposing the gel under UV light and was photographed with a digital UV camera connected together with the visualisation unit (AlphaInnotech). The size of the amplicon were measured by visual comparison to the 1kb DNA marker (Bioline). Phylogenetic typing analysis were carried on the basis of the presence or absence of an amplicon sized 279bp, 211bp and 152bp which belong to chuaA, yjaA and tspE4.c2 genes respectively. 2.4.  Pulse- field gel electrophoresis (PFGE) 2.4.1.  DNA extraction Each isolates was inoculated into 5ml Mueller Hinton Broth and incubated overnight at 37 ºC with gentle agitation. Cells were then harvested by placing 1ml of culture into 1.5ml microcentrifuge tube and was centrifuged at 13, 000 rpm for one minutes. The supernatant was discarded and the process was repeated until all the 5ml of culture finished. The supernatant was again discarded and pellet of cells was resuspended in 500 µl of 0.5M EDTA buffer (see appendix) and was centrifuged at 13, 000rpm for one minutes to removes broth debris that might be interfering with the extraction processes. The washing step was repeated twice to ensure complete removal of debris. The supernatant was discarded once again and pellet was resuspended in 500 µl of suspension buffer. 2.4.2.  Preparation of low melting point (LMP) agarose To prepare the LMP agarose, 3g of SeaKem PFGE agarose (BioRad) were dispensed into 100ml of TE buffer (see appendix) in a universal bottle. It were then heated to dissolve. Agarose was transferred to a 56 ºC waterbath until needed. 2.4.3.  Preparation of the bacterial plugs The wells of PFGE plug molds were numbered. 3 plugs was prepared for each isolates. Then, 750 µl of LMP agarose was added immediately into each cell- buffer suspension and carefully mixed by pipetting up and down several times and be careful not to induce any formation of bubbles. The mixture of cells and agarose was quickly pipetted into the well of a plastic PFGE plug molds (BioRad). The wells was filled to the rim and plugs were allowed to solidify at room temperature or chilled for 5 minutes in the refrigerator. 2.4.4.  Lysis of the cells The cells were lysed by adding a mixture of 1ml of proteolysis buffer with 10 µl of Proteinase K stock solution (50mg/ml) (see appendix) into a 1.5ml new labelled microcentrifuge tube. The plugs were removed from the plug molds by peeling the sealant tape below the wells until all tape was removed. The PFGE plastic arm was used to push the plugs out of the molds into the microcentrifuge containing the mix of proteolysis buffer-proteinase K solution. All plugs for one isolates were transferred into the same tubes. Care was taken while pushing the plugs out of the molds as not to tear the fragile plugs. Tubes was then incubated in a heating block at 50 ºC for 24 hours for digestion to take place. 2.4.5.  Washing of the plugs After completion of an overnight incubation, the proteolysis buffer and Proteinase K activity were eliminated by carefully pipetting out the volume, care taken not to tear the plugs. The plugs were then washed with TE buffer. The washing steps was repeated three times, for every half an hour and were held at room temperature to equilibrate the plugs. 2.4.6.  Restriction enzyme digestion After completion of the washing steps, wash buffer was removed in the final wash leaving only agarose gel in the tubes. Then, 300 µl of 1X restriction enzyme buffer specific to the enzyme used was pipetted in each tubes containing the agarose plugs and was let to equilibrate at room temperature for 10 minutes. The restriction buffer was then discarded, taking care not to tear the plugs. Next, 300 µl of restriction buffer containing 50U of Xbal enzyme was added into the tubes and was incubated in an incubator for 24 hours at 37 ºC specific to the optimal temperature for Xbal enzyme. 2.4.7.  Pulse- field gel electrophoresis 2.4.7.1.  Electrophoresis gel preparation. After incubation, restriction enzyme reaction was stopped by addition of 200 µl of 50mM EDTA. Plugs were cooled at 4 ºC until needed. Then, a (1%) agarose gel was prepared by heated to dissolved 3g of PFGE grade agarose (BioRad) into 300ml of 0.5X TBE buffer over magnetic hot plate with constant stirring or in the microwave and swirl to dissolved. The agarose was then poured into a casting tray that has been placed with PFGE comb and let to solidify at room temperature. The enzyme- buffer was aspirated and one plug of each isolates was loaded into the gel. Care was taken not to tear the plugs. Then, a thin slice high range and mid- range lambda molecular weight marker (New England Biolabs) was loaded into the wells next to each other. After all samples was loaded into wells, the wells were sealed with melted LMP agarose. 2.4.7.2.  Electrophoresis Run The electrophoresis was performed by using a CHEF mapper (BioRad) which subsequently was filled with approximately 3 liters of 0.5ml TBE buffer. The running buffer was let to cool approximately at 14 ºC before turning on the pump. The run time was set for 24 hours at 6 Vcm-1 with 120 º angle using switch time of 2.16 sec to 54.17 sec. 2.4.7.3.  Gel staining Once the run was complete, the gel was stained with 3X Gel red nucleic acid stain (Biotium) with approximately 200ml distilled water and was gently agitated on rotary shaker for 20 minutes. The gel was then visualised under UV transilluminator and a picture was taken once optimal image obtained.

Mans Relationship to the Land in John Steinbecks Grapes of Wrath Essa

     Ã‚  Ã‚  Ã‚  Ã‚   Man's relationship to the land undergoes a transformation throughout John Steinbeck's Grapes of Wrath. Initially, back in Oklahoma, each family feels a strong attachment to the land because the ancestors of these farmers fought and cleared the Indians out of the land, made it suitable for farming, and worked year after year in the fields so that each generation would be provided for. Passing down the land to successive generations, the farmers come to realize that the land is all that they own. It is their family's source of sustenance. However, the strong bond between man and the land is broken when the bank comes to vacate the tenants during hard times.   The tractors hired by the bank literally tear down the bond between man and the land. Due to the eviction, the farmers are forced to move to California, where work is supposedly in demand. As each family takes off for California, it no longer feels a connection to the lands through which it is traveling. Once it reaches California, it feels no connection to its land. For the first time, it is forced to be dependent on somebody else's generosity in distributing jobs, and most importantly, somebody else's land. Thus, in California, the relationship between man and land is not as strong as it was in Arkansas and Oklahoma. The change in this relationship is due in part to the mercilessness of the bank, and in the end, man loses because its connection to the only significant thing it has ever owned is gone. Once the families travel to California, each family member's soul stays back in Oklahoma, making it difficult to adjust to working on lands that have not been cultivate d by their own family for generations.    The land of each generatio... ...job, but instead, little is offered, because of the numbers that they are coming in. Ultimately, one must conclude that no matter how poor a family may be, without land, all is lost in pursuit of a replacement of the heritage that has been destroyed by a superior power. Works Cited and Consulted: Conder, John J. "Steinbeck and Nature's Self: The Grapes of Wrath." John Steinbeck, Modern Critical Views. New York: Chelsea House Publishers, 1987. 125-140. French, Warren. John Steinbeck. Boston: Twayne Publishers, 1975. Levant, Howard. "The Fully Matured Art: The Grapes of Wrath." John Steinbeck, Modern Critical Views. New York: Chelsea House Publishers, 1987. 35-62. Steinbeck, John. The Grapes of Wrath. New York: Penguin Books, 1978. Wallsten, Robert and Steinbeck, Elaine. Steinbeck: A Life in Letters. New York: The Viking Press, 1975.

The Immortal Life of Henrietta Lacks by Rebecca Skloot Essay -- Medica

After her death in 1951, for six decades, Henrietta Lacks did not exist in the eyes of the society, but her cells did. How? Well, the answer is quite simple. HeLa Cells are the first immortal human cells. These cells never die and multiply every twenty-four hours. After spending 10 years to perfect her first book, author of The Immortal Life of Henrietta Lacks, Rebecca Skloot essentially captured the life, the death, and aftermath of Henrietta Lacks’ life. With controversial issues regarding science, ethics, race, and class Skloot takes us on an extraordinary journey. From the â€Å"colored† ward of Johns Hopkins Hospital in the 1950s to stark white laboratories with freezers full of HeLa cells, from Henrietta’s small, dying hometown of Clover, Virginia to East Baltimore, where her children and grandchildren live and struggle with the legacy of her cells, Skloot remarkably shows the story of the Lacks family is inextricably connected to the dark history of experi mentation on African Americans along with the issue of bioethics, and legal battles over whether we control the stuff we are made of. The most intriguing aspect of this story is how is it that HeLa cells were used to develop the polio vaccine, uncover secrets of cancer, viruses’, and the effects of the atomic bomb, and help lead to important advancements for vitro fertilization, cloning, and genes mapping, yet, her five children are not even covered by medical insurance. Can’t the family sue for a profit? This question has been asked multiple times and in various forms, but the answer remains controversial. As Skloot addresses in her book, many lawyers point out that the family â€Å"cannot sue over the cells being taken†¦[but] they could attempt to stop HeLa research through a law... ... May 2010. Moreno, Jonathan D. "Lessons Learned A Half-Century of Experimenting on Humans." The Humanist Sept. 1999: 9. Questia. Web. 31 May 2010. "Nazi Neighbour; Nathan Gasch Moved to the US to Escape His Holocaust Memories but Six Decades on He Discovered the Man Next Door Was an SS Guard. at the Camp Where He Had Been a Prisoner." The Mirror (London, England) 6 Oct. 2007: 31. Questia. Web. 31 May 2010. "S. Fla. Hospital Called 'Most Dangerous' - Health News Story - WPLG Miami." Just News | Miami News, Fort Lauderdale News, Florida News, Weather | WPLG Local 10. Local 10 News, 14 Sept. 2009. Web. 27 May 2010. . Skloot, Rebecca. The Immortal Life of Henrietta Lacks. New York: Crown, 2010. Print. Williams, Patricia J. "State of Denial." The Nation 13 Oct. 2003: 10. Questia. Web. 31 May 2010.